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Cellular and subcellular study of the in vitro immune response

1971; Wiley; Volume: 1; Issue: 2 Linguagem: Inglês

10.1002/eji.1830010207

1521-4141

J Panijel, Françoise Delamette, Micheline Leneveu,

Immunotherapy and Immune Responses

Abstract A culture system using dissociated spleen cell suspensions and phage Φ X174 as antigen has been described. In this system, a cell suspension, originating either from normal (not intentionally immunized) rabbit or from germ‐free mice may be processed as a responder unit: after first addition of Φ X174, a specific reaction has been obtained which presented the various characteristics of a primary response, with respect to size, kinetics, antigen dose, length of stimulation period and 19 S nature of antibodies; in addition, this in vitro primary response was affected by actinomycin and did not involve any increase in the count of viable cells nor in [ 3 H] thymidine incorporation. Priming of the suspension without concomitant antibody production has been also obtained. After this primary response, a second addition of Φ X provoked a new reaction which behaved as a secondary response, with respect to various properties, particularly: amount and 7 S nature of the antibodies, insensitivity towards actinomycin, strong enhancement in the number of viable cells and [ 3 H]‐thymidine incorporation. A true 19 S memory was demonstrated, the expression of which was limited to the early phase of the secondary response. Besides cellular data, subcellular analysis of the primary response showed a sequence of events similar to those in materials originating from immunized animals, that is to say: early appearance, at the level of perinuclear zone, of antibodies having properties of subunits of IgM, silent period during which neither perinuclear nor microsomal activity could be detected, and, preceding the secretion in the culture medium, appearance of 19 S IgM at the level of microsomes. These results show that the same model for explaining the intra‐cellular course of the in in vivo primary response is also relevant for the in vitro primary response.